摘要
目的观察现今临床常用的环孢素（CsA）、他克莫司（FK506）、骁悉（MMF）和雷帕霉素（Rap）4种免疫抑制剂,对小鼠骨髓来源的树突细胞（DC）分化、成熟和功能的影响,探讨这些药物是否具有诱导免疫耐受的可能性。方法在小鼠骨髓来源的DCp培养过程中加入粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白介素-4（IL-4）和四种不同药物,用流式细胞仪检测各组DC表面CD11c、CD40和CD80的表达。MTT比色法行体外混合淋巴细胞反应（MLR）观察每组对同种T细胞的刺激增殖能力。结果各药物处理组与对照组形态上无差别（P〉0.05）,CD11c表达都在80﹪以上,经LPS刺激后CSA组和FK506组细胞表型分化成熟（CD11c＋,CD40＋,CD80＋）,而MMF组和RAP组无明显变化（CD11c＋,CD40-,CD80）,各处理组对同种异体T细胞增殖抑制程度顺序为：RAP〉MMF〉FK506〉CSA。结论通过对这些经典的免疫抑制药物对DC影响的研究,为在临床上合理的选择免疫抑制药物及实验室中制备致耐性树突细胞（Tol-DC）提供理论根据。
Objective Toinvestigatetheeffectof Rapamycin（RAP）,FK506, Cyclosporin A（CSA）and mycophenolate mofetil（MMF）on the differentiation, maturation and function of murine bone marrow derived dendritic cells（DC）in vitro and to explore the possibility of tolerance induction.Methods DC progenitors were propagated from mouse bone marrow with granulocyte-macrophage colony-stimulating factor（GM-CSF）plus IL-4 for 6 days in the presence or absence of CSA（1.0μg/ml）,FK506（10ng/ml）,RAP（20ng/ml）and MMF（0.1μm）respectively.During DC culture, morphology of the cells was observed under electromicroscopy.Cell surface expression of CD11c,CD40 and CD80 was analyzed by flow cytometry.The antigen-presenting function of DC was determined in one-way mixed leukyocyte reactions.Results Addition of CSA,FK506,MMF or RAP to the culture did not change DC appearance. Exposure of DCs during maturation to CSA or FK506 resulted in the expression of DC phenotypic markers,including CD40 and CD80,as determined by flow cytometry.But DC differentiated in the presence of RAP or MMF had decreased expression of CD40 and CD80 compared with control-DC.All immunosuppressive agents significantly reduced the allostimulatory ability of DCs as measured by the primary mixed lymphocyte reaction（RAPMMFFK506CSA）.Conclusion Our results demonstr- ate different effects of RAP and calcineurin inhibitors on Ag presentation.Selection or combination of immunosuppressive agents for clinical application should take these effects into consideration.Whether these immunosuppressive agents can be used for the generation of tolerogenic DCs deserves further study.
关键词
树突细胞
免疫抑制剂
移植耐受
Dendritic cells Immunosuppressive drugs Immune tolerance
